Cell-Free DNA, High-Mobility Group Box-1, and Procalcitonin Concentrations in Dogs With Gastric Dilatation–Volvulus Syndrome

Abstract

Canine gastric dilatation–volvulus (GDV) is a life-threatening disease characterized by extensive tissue ischemia, tissue hypoperfusion, and systemic inflammation. Biomarkers that better reflect the severity of gastric necrosis and systemic inflammation would aid clinicians in the management of these patients. This study aimed to investigate the prognostic significance of cell-free DNA (cfDNA), high-mobility group box-1 (HMGB1), and procalcitonin (PCT) in dogs with GDV. Concentrations of cfDNA, HMGB1, and PCT were measured in citrated plasma samples collected from 29 dogs with GDV at hospital admission. Additional data collected included baseline lactate concentrations, APPLEfast score, evidence of gastric necrosis, occurrence of postoperative complications, and outcome. Twenty-four healthy dogs were sampled as controls. Continuous variables between groups were compared with the Mann–Whitney U and correlations between continuous variables were assessed by calculation of Spearman’s correlation coefficient. Alpha was set at 0.05. Dogs with GDV had significantly greater concentrations of cfDNA, HMGB1, and PCT compared to controls (P = 0.0009, P = 0.004, and P = 0.009, respectively). PCT concentrations were significantly higher in non-survivors compared to survivors (P = 0.008). Dogs with gastric necrosis had significantly greater lactate concentrations compared to dogs without gastric necrosis (P = 0.0005). The APPLEfast score was not prognostic. Lactate and PCT concentrations were moderately, positively correlated (rs 0.51, P = 0.0005). Concentrations of the inflammatory biomarkers cfDNA, HMGB1, and PCT are increased in canine GDV. Only lactate and PCT concentrations were prognostic in this population of GDV dogs and were predictive of the presence of gastric necrosis and of non-survival to hospital discharge, respectively.

Effect of sample type on plasma concentrations of cell‐free DNA and nucleosomes in dogs

“…The final diagnoses for most dogs were not known at the time of recruitment and hence the population enrolled contained more dogs with neoplastic diseases than anticipated. The overall cfDNA and nucleosome concentrations were lower than reported for other patient populations,19 20 22 perhaps reflecting a lower disease severity in this cohort compared with others. Despite this, the cfDNA and nucleosome concentrations covered a wide range (~twofold for cfDNA and ~sevenfold for nucleosomes) and provided a solid dataset to assess for evidence of proportional biases between sample types.…”

“…Concentrations of both biomarkers were measured contemporaneously. Concentrations of cfDNA were measured in triplicate using a benchtop fluorimeter (Qubit 3.0 Fluorometer, Life Technologies, Waltham, MA, USA) and relevant reagents (Quant‐It HS dsDNA Kit, Life Technologies, Waltham, MA, USA) according to the manufacturers’ instructions and as previously described 19 22. Mean values of the three measurements were used for subsequent analyses.…”

“…In people, concentrations of cfDNA are also increased after myocardial infarction and trauma 17 18. Increased cfDNA concentrations have been identified in dogs with sepsis,19 trauma,20 neoplasia,21 gastric dilation volvulus (GDV) syndrome22 and immune-mediated haemolytic anaemia (IMHA) 23 24…”

Neutrophil-Extracellular Traps, Cell-Free DNA, and Immunothrombosis in Companion Animals: A Review

“…Increased concentrations or gene expression of inducers of NETosis (eg, IL-8, TNF-α, and HMGB1) is reported in veterinary patients with a variety of clinical conditions. 56,62,70,200,202,212 This suggests NETosis may occur in various disease states, but it should be recognized that these markers are not specific for NETosis but contribute to numerous inflammatory and cell death pathways. 28,33,163 As discussed above, NETosis is an umbrella term for a variety of molecular pathways that result in DNA release, 94 so it is possible that NETs may be produced without concurrent increases in the concentrations of a particular inducer.…”

High Mobility Group Box-1 and Pro-inflammatory Cytokines Are Increased in Dogs After Trauma but Do Not Predict Survival

“…The half-life of HMGB-1 is ~11 h in people ( 16 ). Concentrations of HMGB-1 are increased in dogs with gastric dilation-volvulus (GDV) ( 17 , 18 ), dogs with sepsis ( 19 ), and correlate with plasma nucleosome concentrations in critically ill horses ( 20 ), suggesting it may provide information about tissue injury in dogs with trauma.…”